LOCUS       sequence-204315-        4440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human eGFP with homology arms for Lamin A/C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4440)
  AUTHORS   Paix A, Folkmann A, Goldman DH, Kulaga H, Grzelak MJ, Rasoloson D, 
            Paidemarry S, Green R, Reed RR, Seydoux G
  TITLE     Precision genome editing using synthesis-dependent repair of 
            Cas9-induced DNA breaks.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114.
  PUBMED    29183983
REFERENCE   2  (bases 1 to 4440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114."
COMMENT     Sequence Label: pAP1716-1
FEATURES             Location/Qualifiers
     source          1..4440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     194..211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     195..211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(756..758,759..1469)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(756..758,759..1469)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(798..819)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1059..1078)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1406..1427
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2035..2051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2035..2051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2048..2070)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2059..2075
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2083..2089,2090..2107,2108..2113))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2128..2149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2266..2283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2437..3025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2517..2536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3196..3987,3988..4056))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3819..3838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4057..4161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4229..4247
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4285..4307)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4407..4426
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
       61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      181 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatcctc
      241 tccctccttg gctctgccct ctagcccaga aggtctgagg caatgggggc aagcttggag
      301 ccgacagtgc tgagcaggca ggagccaaga gaggggaagc ttgagcctca cgcagttagg
      361 ggtgcgctgg agagggtggg gcccgactcc gccacacccc aacggtcctt ccccctcctc
      421 accactcccg cccccacccc caatggatct gggactgccc ctttaagagt agtggcccct
      481 cctcccttca gaggaggacc tattagagcc tttgccccgg cgtcggtgac tcagtgttcg
      541 cgggagcgcc gcacctacac cagccaaccc agatcccgag gtccgacagc gcccggccca
      601 gatccccacg cctgccagga gcaagccgag agccagccgg ccggcgcact ccgactccga
      661 gcagtctctg tccttcgacc cgagccccgc gccctttccg ggacccctgc cccgcgggca
      721 gcgctgccaa cctgccggcc atggagaccc cgtccgtgag caagggcgag gagctgttca
      781 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      841 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      901 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      961 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1021 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1081 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1141 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1201 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1261 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1321 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1381 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1441 tcactctcgg catggacgag ctgtacaagc agcggcgcgc cacccgcagc ggggcgcagg
     1501 ccagctccac tccgctgtcg cccacccgca tcacccggct gcaggagaag gaggacctgc
     1561 aggagctcaa tgatcgcttg gcggtctaca tcgaccgtgt gcgctcgctg gaaacggaga
     1621 acgcagggct gcgccttcgc atcaccgagt ctgaagaggt ggtcagccgc gaggtgtccg
     1681 gcatcaaggc cgcctacgag gccgagctcg gggatgcccg caagaccctt gactcagtag
     1741 ccaaggagcg cgcccgcctg cagctggagc tgagcaaagt gcgtgaggag tttaaggagc
     1801 tgaaagcgcg gtgagttcgc ccaggtggct gcgtgcctgg cggggagtgg agagggcggc
     1861 gggccggcgc ccctggccgg ccgcaggaag ggagtgagag ggcctggagg ccgataactt
     1921 tgccatagtc tcctccctcc ccggaactgc ccccagcggg tgactggcag tgtcaagggg
     1981 aattgtcgat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     2041 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2101 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2161 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2221 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2281 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3181 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3481 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3781 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4201 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4261 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4321 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4381 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
//
